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Biomedical subjects

J R Wolff

Publications and source records attributed to J R Wolff.

At least 73 records · Page 4Linked to original sources
PubMed1991

Are there cholinergic through-fibers in the superior cervical ganglion of the mouse?

Choline acetyltransferase immunocytochemistry was used to detect the presence and distribution of cholinergic through-fibers in the superior cervical ganglion of adult mice. The results revealed a great number of choline acetyltransferase-positive axons in the cervical sympathetic trunk and the varicose terminal axons involved in the innervation of the principal ganglionic cells within the ganglion. Immunostained axons were scarce or absent in the external or internal carotid nerves. The immunocytochemical results argue against the physiological importance of cholinergic through-fibers in the postganglionic nerves.

Animals↗
PubMed1991

Cholinergic innervation of the mouse superior cervical ganglion: light- and electron-microscopic immunocytochemistry for choline acetyltransferase.

The cholinergic innervation of the mouse superior cervical ganglion was investigated by means of immunocytochemistry using a well-characterized monoclonal antibody against choline acetyltransferase (ChAT). Immunopositive nerve fibers entered the superior cervical ganglion from the cervical sympathetic trunk. Light-microscopically, these fibers appeared to be heterogeneously distributed among the principal ganglion cells. The rostral part of the ganglion contained more ChAT-positive fibers then the middle or the caudal one. The axons branched several times before forming numerous varicosities. Most of the ChAT-stained fibers and varicosities aggregated in glomerula-like neuropil structures that were surrounded by principal ganglion cell bodies, whereas others were isolated or formed little bundles among principle neurons. None of the neurons or other cell types in the ganglion exhibited ChAT-positivity. ChAT-immunoreactive fibers disappeared from the ganglion 5 or 13 days after transection of the cervical sympathetic trunk. At the ultrastructural level, most axon terminals and synapses showed ChAT-immunoreactivity. An ultrastructural analysis indicated that immunostained synapses occurred directly on the surface of neuronal soma (1.8%) and dendritic shafts (17.6%). Synapses were often seen on soma spines (18.4%) and on dendritic spines (62.2%). All immunoreactive synapses were of the asymmetric type. The results provide immunocytochemical evidence for a heterogeneous cholinergic innervation of the ganglion and the principal neurons.

Animals↗
PubMed1991

Morphology of neurons in the rat basal forebrain nuclei: comparison between NADPH-diaphorase histochemistry and immunohistochemistry of glutamic acid decarboxylase, choline acetyltransferase, somatostatin and parvalbumin.

Transversal sections through the basal forebrain of 11 adult male rats were immunostained for glutamic acid decarboxylase (GAD), choline acetyltransferase (ChAT), somatostatin (SOM) and parvalbumin (PARV). Immunohistochemistry of ChAT, PARV, and SOM was combined with histochemistry of NADPH-diaphorase (NADPH-d) to obtain information on the colocalization of various neuroactive substances and this enzyme and to facilitate the recognition of morphological details of double-stained neurons. The distribution patterns of GAD- and PARV-immunoreactive cells were only in part congruent in basal forebrain nuclei in the rat. In the medial septal nucleus (MS) and the vertical limb of the diagonal band (vDB) PARV-immunopositive neurons were homogeneously scattered inside the nucleus, whereas the GAD-immunoreactive cells were much more numerous in the lateral part of this nuclear complex. In the horizontal limb of the diagonal band (hDB) and the nucleus preopticus magnocellularis (NPM), where GAD-immunoreactive cells occurred in high number, only very few cells contained PARV-immunoreaction product. In the substantia innominata-nucleus basalis Meynert complex (SI-NB) and in the ventral pallidum (VP) the neuropil was heavily stained with the GAD-immunoreaction product. The number of GAD-positive cells appeared low in the SI-NB, but much higher in the VP. In this nucleus GAD- and PARV-immunoreactive cells seem to be identical. PARV-positive neurons are very sparse in the SI-NB. Double-staining of PARV-immunoreactivity and NADPH-d was not registered. These nuclei were the only ones in which some cells with SOM-like immunoreactivity were observed. Among ChAT-positive neurons those double-stained with NADPH-d occurred in moderate number, but with obvious regional differences. In MS-vDB and the marginal zone of hDB the two neuron groups were intermingled, but only in the innermost part of the hDB ChAT-single-immunostained cells form aggregates, which were also typical of the zone in the SI-NB that surrounds and infiltrates the globus pallidus (GP). Double-labelled cells were more frequent in the lateral aspect of the NPM and SI-NB. Cells single-stained for NADPH-d were frequent in the MS-vDB along the border toward the lateral septal nuclei, but low in number in the NPM, VP and SI-NB. The functional aspects of the occurrence of GAD-immunoreactive cell aggregates in the lateral preoptic area (LP) and the lateral hypothalamic area (LH) were discussed with special regards to extrinsic GABAergic input in the dorsal SI-NB.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1990

Vinpocetin protects against excitotoxic cell death in primary cultures of rat cerebral cortex.

The protective effect of vinpocetin, a drug clinically useful in brain hypoxia/ischemia, was examined in vitro on cerebrocortical cultures treated with glutamate and related excitotoxins. The extent of cell death was quantified by measuring lactic dehydrogenase activity released from damaged cells into the culture medium. Vinpocetin partially protected the cortical cells against cell death induced by N-methyl-D-aspartate, quisqualate and kainate, indicating that the drug exerts a direct protective action on cerebrocortical cells bearing excitatory amino acid receptors.

Amino Acids↗
PubMed1990

Lack of excitotoxic cell death in serum-free cultures of rat cerebral cortex.

The excitotoxicity of glutamate, N-methyl-D-aspartate (NMDA), quisqualate and kainate were compared in rat cerebro-cortical cell cultures grown in different media. Excitotoxic cell death was examined by phase contrast microscopy and by the measurement of lactic dehydrogenase (LDH) activity released in the culture medium. Cells grown for 14 days in a serum-free, defined (N2) medium were resistant, while sister cultures maintained in the presence of serum responded to the excitotoxins by a significant degree of cell death. Cells cultured in serum-free medium for 7 days, and fed by a serum-containing one for an additional week exhibited vulnerability to the excitotoxins. It is concluded that the development of excitotoxicity in rat cortical cultures depends on one or more serum constituents.

Amino Acids↗
PubMed1990

Golgi-like demonstration of "dark" neurons with an argyrophil III method for experimental neuropathology.

A silver method is proposed for the selective, well-contrasted and reproducible demonstration of "dark" neurons in frozen, vibratome and paraffin sections cut at a thickness of 5 to 200 microns from aldehyde-fixed brains. The Golgi-like staining of the dendrites enables assorting of "dark" neurons according to characteristic neuron classifications. The staining procedure includes an esterification with 1-propanol, a treatment with diluted acetic acid and development. The esterification strongly increases the argyrophilia of both "dark" neurons and mitochondria. Unwanted co-staining of mitochondria is suppressed by the acetic acid treatment, while a special developer is used to render the staining controllable. The applicability of the method to experimental neuropathology is demonstrated by Golgi-like staining of "dark" neurons in rat brains exposed, before transcardial perfusion-fixation and delayed autopsy, to various pathological conditions including ischemia, hypoglycemia, trauma, status epilepticus, deafferentation and poisoning with kainic acid, colchicine and sodium azide, respectively.

Animals↗
PubMed1990

Structures with GABA-like and GAD-like immunoreactivity in the cervical sympathetic ganglion complex of adult rats.

The distribution of gamma-aminobutyric acid (GABA)-like and glutamate decarboxylase (GAD)-like immunoreactivity was studied in the cervical sympathetic ganglion complex of rats, including the intermediate and inferior cervical ganglia and the uppermost thoracic ganglion. GABA-positive axons may enter the ganglion complex via its caudal end. Others apparently arise from small GABA-positive cell bodies which are scattered among principal neurons, within clusters of SIF cells and in bundles of GABA-negative axons. The majority of these cells is located in the lower half of the ganglion complex. Principal neurons did not react with antibodies against GABA or GAD. An unevenly distributed mesh-work of GABA-immunoreactive axons was seen in each of the ganglia. Immunoreactive axons formed numerous varicosities. Some of them were aggregated in a basket-like form around a subpopulation of GABA-negative principal ganglion cell bodies. Electron-microscopic immunocytochemistry revealed that GABA-positive nerve fibers establish asymmetric synaptic junctions with dendritic and somatic spines of principal neurons, whereas postsynaptic densities are inconspicuous or absent on dendritic shafts and somata. The results suggest that in the cervical sympathetic ganglion complex principal neurons are not GABAergic, but are innervated by axons which react with both antibodies against GAD and/or GABA antibodies and originate from a subpopulation of small neurons.

Animals↗
PubMed1990

An unbiased estimation of the total number of synapses in the superior cervical ganglion of adult rats established by the disector method. Lack of change after long-lasting sodium bromide administration.

Previous physiological and morphological studies suggested that sodium bromide promotes synaptogenesis of implanted cholinergic nerves in the superior cervical ganglion of adult rats. To check whether sodium bromide also modifies synaptic numbers in the intact ganglion, quantitative electron microscopy was used to determine the total number of synaptic junctions in the superior cervical ganglion of adult rats. Untreated controls were compared with animals which drank water containing 280 mg ml-1 sodium bromide for 7 days. The disector method, an unbiased estimator of volume density of certain particles, has been adapted to this particular case. To accomplish the task, an on-line counting procedure was developed, which permitted the efficient adaptation of the disector method for the superior cervical ganglion, in which the synapses are known to be distributed sparsely. Three pairs of (control and treated) ganglia have been completely processed by three independent examiners. The estimated number of synapses in the ganglia ranged from 4 to 8 million while the volumes of the ganglia varied from 0.65 to 0.90 mm3. Evaluation of the results showed that variations in the total number of synapses were in each case proportional to differences in ganglionic volumes. This suggests that: (1) sodium bromide does not lead to changes in density of intrinsic synapses; and (2) the morphogenetic action of sodium bromide on principal ganglion cells previously described is essentially postsynaptic and requires additional presynaptic elements to increase the number of synapses.

Animals↗
PubMed1990

An argyrophil-III method for the selective demonstration of mitochondria in aldehyde-fixed rat brain.

Based on recent achievements in physicochemical aspects of histological silver staining, a selective and reliable method was elaborated for the demonstration of mitochondria in frozen sections of aldehyde-fixed rat brain. Areas of similar architecture were stained with comparable intensity both within one section or in serial sections. The distribution patterns of mitochondria can be examined at low magnification or even with the unaided eye. At high magnifications, rounded and elongated profiles with the size and shape of mitochondria clearly stand out against a pale background. Electron microscopy revealed a high degree of selectivity and completeness of the mitochondrial staining.

Aldehydes↗
PubMed1990

Sodium bromide treatment influences the plasticity of somatosensory responses in the rat cortex as induced by enucleation.

Effects of sodium bromide were studied on central neuroplasticity induced by early binocular enucleation. It has previously been found that enucleation on the day of birth, but not later than the first postnatal week, resulted in changes in the occipital cortex, such as the invasion of somatosensory evoked activity into the visual cortex areas. The present results showed that sodium bromide treatment extended at least up to 15 days after birth, the critical period during which somatosensory projections could be modified by visual deafferentation. Together with observations of Frost [J. comp. Neurol. (1981) 203, 227-256; Devl Brain Res. (1982) 3, 627-636], the present results suggest a mutual dependency of visual and somatosensory projection development. The present study is the first demonstration that the critical period of development, during which a specific type of neural plasticity can be induced, may be prolonged by pharmacological means, i.e. by chronic treatment with sodium bromide.

Animals↗
PubMed1990

Fine structural changes in the superior cervical ganglion of adult rats after long-term administration of baclofen, a GABAB receptor agonist.

The aim of the present study was to examine the fine structural changes in the superior cervical ganglion of adult rats following baclofen treatment. On the addition of this GABAB receptor agonist there was a transient increase in the number of coated vesicles and pits in the dendrites of principal sympathetic neurons. As compared to the control ganglia, we found a significantly higher number of synapses which had coated pits attached to the postsynaptic densities. In interpreting these observations it is suggested that the GABAB receptors play some role in mediating the synaptogenetic action of GABA, although the formation of vacant postsynaptic densities and spines was not seen after baclofen treatment.

Action Potentials↗
PubMed1990

Cerebral endothelial cell-derived laminin promotes the outgrowth of neurites in CNS neuronal cultures.

The aim of the present study was to elucidate whether the endothelial cells, besides providing the structural basis for the blood-brain barrier (M. Bradbury, Concept for a Blood-Brain Barrier, 1979; F. Joó, Nature 321, 197-198, 1986) can also produce laminin and, via its regulate the growth of neurites of neurons originating from the central nervous system (CNS). In immunohistochemical investigations, the presence of laminin-like immunoreactivity could be detected in the cytoplasm and on the surface of endothelial cells derived from brain microvessels. Further, the neurite-promoting effect of laminin on CNS neurons was also noted in a co-culture system designed especially for conditioning nerve cell cultures continuously with materials released from the cerebral endothelial cells. It is suggested that the endothelial cells may be involved via their laminin secretion in the regulation of axonal growth and guidance in the CNS.

Animals↗
PubMed1990

Postnatal development of the excitatory amino acid system in visual cortex of the rat. Changes in ligand binding to NMDA, quisqualate and kainate receptors.

The postnatal development of the ligand binding to N-methyl-D-aspartate (NMDA), quisqualate and kainate receptor sites was examined in whole homogenates of the visual cortex of rats, aged 2-360 days. As selective ligands, [3H]CPP (3-(2-carboxypyperazine-4-yl)-propyl-1-phosphonic acid, [3H]AMPA (RS-alpha-amino-3-hydroxy-5-methyl-isoxazole-4-propionic acid) and [3H]KA (kainic acid) were used, respectively. The binding of CPP was low in newborns, rapidly increased from the second postnatal week, reached its maximum between weeks 2 and 3, then slowly declined up to the age of 1 year. In contrast, the binding of AMPA and kainate was high perinatally, increased rapidly up to day 6 after birth to reach an early maximum value, then gradually decreased to adult values which were attained at an age of 3-4 weeks. These age-related changes were derived from alterations in the density of binding sites, which, in the case of AMPA, was accompanied by an increase in binding affinity. The results, compared with the developmental time-course of excitatory synapses, indicate that, in the immature cerebral cortex, NMDA receptors may be primarily involved in synaptic transmission, whereas quisqualate and kainate receptors may play some other (e.g. trophic) roles.

Aging↗
PubMed1990

Postnatal development of the excitatory amino acid system in visual cortex of the rat. Changes in uptake and levels of aspartate and glutamate.

The postnatal changes of aspartate (Asp) and glutamate (Glu) levels, as well as the development of the high-affinity uptake of [3H]D-Asp were examined in the visual cortex of rats, aged 2-360 days. Perinatal (day 2) levels of Asp were relatively lower than those of Glu, i.e. about 30 and 60% of the respective values for adults (day 180). Both Asp and Glu levels showed a rapid increase during the second and third postnatal weeks. Asp reached its maximal concentration (day 20) a few days earlier than Glu (day 25). Thereafter, both Asp and Glu levels slowly declined to the adult values which were reached by day 90 and remained unchanged up to day 360. The uptake of D-Asp was low in newborns (5% of that in adult) and progressively increased with age up to about day 20, when adult values were attained. The increase of Asp and Glu levels showed a close correlation with the maturation of uptake capacity. Our findings indicate that the developmental increase of Asp and Glu levels is associated with the maturation of excitatory innervation in the visual cortex, but they do not support the concept of a transient Asp/Gluergic innervation of the immature visual cortex.

Aging↗
PubMed1990

x-y-recording in transmission electron microscopy. A versatile and inexpensive interface to personal computers with application to stereology.

An interface for IBM XT/AT-compatible computers is described which has been designed to read the actual specimen stage position of electron microscopes. The complete system consists of (i) optical incremental encoders attached to the x- and y-stage drivers of the microscope, (ii) two keypads for operator input, (iii) an interface card fitted to the bus of the personal computer, (iv) a standard configuration IBM XT (or compatible) personal computer optionally equipped with a (v) HP Graphic Language controllable colour plotter. The small size of the encoders and their connection to the stage drivers by simple ribbed belts allows an easy adaptation of the system to most electron microscopes. Operation of the interface card itself is supported by any high-level language available for personal computers. By the modular concept of these languages, the system can be customized to various applications, and no computer expertise is needed for actual operation. The present configuration offers an inexpensive attachment, which covers a wide range of applications from a simple notebook to high-resolution (200-nm) mapping of tissue. Since section coordinates can be processed in real-time, stereological estimations can be derived directly "on microscope". This is exemplified by an application in which particle numbers were determined by the disector method.

Animals↗
PubMed1989

Transient increase in ligand binding to quisqualate and kainate sites in cerebral cortex of immature rats.

The postnatal changes in the specific binding of [3H]kainate and [3H]AMPA (RS-alpha-amino-3-hydroxy-5-methyl-4-isoxasolopropionic acid, an agonist of quisqualate receptors) were studied in cerebral cortex of rats, aged 2-360 days. The binding of the two ligands was assayed in whole-tissue homogenates. Similar developmental time courses were found for kainate and AMPA binding, characterized by high perinatal levels, a further increase during the first few days after birth, an early maximum value around the age of one week, and a gradual decrease to adult values which were attained at an age of 3-4 weeks. As revealed by Scatchard analysis, the transient elevation of ligand binding was derived from an increased density of binding sites, which, in the case of AMPA, was accompanied also by an increase in binding affinity. The results indicate that, in the immature cerebral cortex, kainate and quisqualate receptors may play a role other than in synaptic transmission.

Aging↗
PubMed1989

GABAA receptors in the rat stomach may mediate mucoprotective effects.

The occurrence and characteristics of binding sites specific for gamma-aminobutyric acid (GABA) and muscimol in the rat stomach were examined by biochemical and autoradiographic techniques, and the effects of GABAergic model compounds on gastric ulceration induced by chemical irritation was studied in intact and unilaterally vagotomized rats. Specific binding sites for [3H]GABA and [3H]muscimol, which showed the characteristics of GABAA receptors, were demonstrated on gastric membranes. Specific muscimol binding sites were found in all regions of the stomach and were present in both the mucosal layer and the remaining tissue of the stomach. Oral pretreatment of the rats with GABA, selective GABAA receptor agonists, or inhibitors of GABA degradation protected the gastric mucosa against the ulcers induced by acidified ethanol (chemical irritant), in both intact and vagotomized rats. These findings are consistent with the view that a subpopulation of GABAA receptors in the rat stomach may mediate the anti-ulcer effect.

Administration, Oral↗